Proteomics in globe artichoke: Protein extraction and sample complexity reduction by PEG fractionation

被引：20

作者：

Acquadro, Alberto ^{[1
]}

Falvo, Sara ^{[1
,2
]}

Mila, Silvia ^{[2
]}

Albo, Alessandra Giuliano ^{[2
]}

Comino, Cinzia ^{[1
]}

Moglia, Andrea ^{[1
]}

Lanteri, Sergio ^{[1
]}

机构：

[1] Univ Turin, DiVaPRA, I-10095 Grugliasco, TO, Italy

[2] Bioind Pk Canavese, Colleretto Giacosa, Italy

来源：

ELECTROPHORESIS | 2009年 / 30卷 / 09期

关键词：

2-DE; Globe artichoke; MS analysis; PEG; Protein extraction; CYNARA-SCOLYMUS L; POLYETHYLENE-GLYCOL FRACTIONATION; IMMOBILIZED PH GRADIENTS; 2-DIMENSIONAL ELECTROPHORESIS; MICROSATELLITE MARKERS; PHENOLIC-COMPOUNDS; ABUNDANT PROTEINS; GENETIC-VARIATION; OXIDATIVE STRESS; VAR; SCOLYMUS;

D O I：

10.1002/elps.200800549

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here, we report the first leaf proteome analysis for globe artichoke. Three protein extraction protocols were tested and a reproducible Mg/NP-40-based method was established. Ribulose-1,5-biphosphate carboxylase-oxygenase (RuBisCO) is a highly abundant leaf protein, and its presence masks co-localizing, less abundant proteins. To remove RuBisCO from the sample, and thereby improve spot resolution, a PEG fractionation approach was elaborated. 2-DE profiles of various PEG fractions showed that the fractionation procedure was successful in excluding most of the RuBisCO, allowing for the detection of many low-abundance proteins. Western blot analysis was able to confirm the reduction in RuBisCO content achieved by PEG fractionation. In all, 841 distinct protein spots were detected, and 40 of these, selected from the RuBisCO region of the 2-DE profile, were successfully identified by MS. A number of homologues of these proteins also co-localize with RuBisCO in Arabidopsis thaliana.

引用

页码：1594 / 1602

页数：9

共 39 条

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